2.9. Analysis of Apoptosis and Measurement of the Mitochondrial Membrane Potential (MMP)

An apoptosis/necrosis detection kit (ab176749, Abcam, MA, USA) was used to simultaneously monitor apoptotic (green), necrotic (red), and healthy (blue) cells in strict accordance with the manufacturer’s instructions. The stained samples were analyzed by flow cytometry (Beckman Coulter, Brea, CA, USA) and fluorescence microscopy (LSM800, Carl Zeiss, Germany).

The MMP was measured using a JC-1 mitochondrial membrane potential assay kit (C2006, Beyotime Biotechnology, Shanghai, China). Cells were incubated with JC-1 solution at 37 °C for 20 min, and imaging was then performed using a high-content imaging system (Harmony, PerkinElmer, Rodgau, Germany). The ratio of red to green fluorescence intensity was used to determine the MMP.